IN-VITRO STUDY ON CLINICAL PREVALENCE AND ANTIMICROBIAL SUSCEPTIBILITY OF CONJUNCTIVA INFECTING STAPHYLOCOCCUS AUREUS IN HUMANS

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1 IN-VITRO STUDY ON CLINICAL PREVALENCE AND ANTIMICROBIAL SUSCEPTIBILITY OF CONJUNCTIVA INFECTING STAPHYLOCOCCUS AUREUS IN HUMANS S. Jeeva 1*, V.R. Jeya Sree 1, T.Selva Mohan 2, N.C.J.Packia Lekshmi 1, and J.Raja Brindha 1 1 Department of Microbiology, Udaya College of Arts and Science, Vellamodi, India 2 Department of Zoology, Raja Doraisingam Government Arts College, Sivagangai, India. jeevarajaseker@gmail.com (Received on Date: 29 th June, 2012 Date of Acceptance: 29 th July, 2012) ABSTRACT The present report deals with our preliminary experiments designed to evaluate the in-vitro effects of various antibiotics on conjunctiva infecting Staphylococcus aureus by employing antibiotic sensitivity test by disc diffusion method as described by (Cruickshank, 1968). This study was undertaken to investigate whether the antibiotic resistance of Staphylococcus aureus varies with sex or age of patients. A total of 50 isolates of Staph. aureus were obtained from pus of conjunctiva infection. Staphylococcus aureus isolates were isolated most frequently in adult and male children when compared with females. Antimicrobial resistance was commonly found for norfloxacin, pefloxacin, and cefazolin. The isolate showed sensitivity against Gatifloxacin, amicacin, ceprofloxin, and chloramphenicol. From this study we conclude that antimicrobial susceptibilities in Staph. aureus often differed with regard to sex and age of the humans. Keywords:Antimicrobial susceptibility, conjunctiva, Staph. aureus, antibiotics Number of References : 22 Number of Tables: 2

2 INTRODUCTION Worldwide population constitute of about 28% of children and infants who are the most susceptible to diseases than adults which is mainly due to under development of immune system, hormonal imbalance, genetic factors, environmental change, water borne and food borne etc (Moorthiet al., 2001). Eye is most interesting organ due to its drug disposition characteristics. For ailments of the eye,topical administration is usually preferred over systemic administration, before reaching theanatomical barrier of the cornea, any drug molecule administered by the ocular route has to crossthe precorneal barriers(patelet al., 2010).Staphylococci are routinely isolated from conjunctiva in clinical practice (Biberstein et al., 984). Besides their role as commensals on mucosal surfaces and the skin, Staphylococci are often involved in a wide variety of diseases in humans (Klooset al., Staphylococcal infections are frequently treated with antibiotics and, consequently, antibiotic resistance and or acquired resistance have developed(jorgensenet al., 1999). Staphylococcus aureus is frequently associated with suppurative infections and is recognized as a resident member of the microflora of the skin of humans and dogs (Kloos, 1990). Staph. aureus has been isolated in very similar percentages from the skin and other sites (Bibersteinet al., 1984). Only a few preliminary studies have been conducted to determine whether the antibiotic susceptibility of a specific bacterium varies with (i) sex and (ii) age of the individual (Flournoyet al., 1989; Flournoyet al., 1979; Hoekstra and Paulton, 1996). The purpose of this study was to examine the isolate, Staph. aureus with particular reference to three factors: (i) the isolated to determine whether the site of isolation influences antimicrobial susceptibility for a specific bacterium (Hoekstra and Paulton, 1996); (ii) the isolated from male and female given the probable influence of sex in terms of virulence and antibiotic sensitivity (Picard and Goullet, 1989)and (iii) the effect of age to determine whether antibiotic resistance correlated with age of the human, as previously claimed. MATERIALS AND METHODS Pus samples were obtained from Bejansingh eye hospital, Nagercoil, Kanyakumari, Tamil Nadu, India. Adult and Children of both sexes were chosen for identification and susceptibility testing over a 6 month period (2011 Jan-June). Patients selected for this study had no known history of prior antibiotic therapy. The bacterial isolates were identified and labelled as to source, male or female, adult or Children. After growth, Staphylococcal isolates were identified according to their characteristics as outlined in Bergey s Manual of Determinative Bacteriology (Euzeby, 1997; Holtet al., 1994) and the Manualof Clinical Microbiology (Murray and Kloos, 1980). Only isolates identified as Staph. aureus was selected for this study. After growth on sheep blood agar,staph. aureus was identified on the basis of colony characteristics, Gram stain, pigment production, acid production on D-mannitol, free coagulase and the slide test for detection of clumping factor. Once samples were identified, the staphylococcal strains were tested for susceptibility to antibiotics by disc agar diffusion aspreviously described (Hoekstra and Paulton, 1996). Discs of antibiotics commonly used in clinical medicine for infective diseases were

3 tested: Gatifloxacin, Amicacin, Norfloxacin, Pefloxacin, cefazolin, Ceprofloxin, Chloromphenicol, Tobramycin, and Moxifloxacin. After measuring the zones of inhibition, the strains were classified as sensitive, intermediate or resistant to the drug according to the literature (Jorgensen et al., 1999). RESULTS AND DISSCUSSION A total of 50 isolates of Staph. aureus was obtained from male and female, adult and children from January-June Table 1 shows the frequency of isolation of these staphylococcal isolates according to the sex and age of the patients. The study included more male (37) than female (23). The isolates from male and female and adult and children is shown in Table 2. Antimicrobial resistance was commonly found for norfloxacin, pefloxacin, and cefazolin. The isolate showed sensitivity against Gatifloxacin, amicacin, ceprofloxin, and chloramphenicol. It is found that the isolates showed moderately sensitive to topramycin and moxifloxacin. Table 1: Frequency and distribution of Staphylococcus aureus isolated from 50 patients (Jan-Nov 2010) Sample Male Adult Male children Female adult Female children Pus from conjunctiva Table 2: Percentage susceptibilities of Staphylococcus aureus isolated from the pus of conjunctiva against various antibiotics. S.no Antibiotics Male Adult Male Female Female children children adult 1 Gatifloxacin S S S S 2 Amicacin S S R S 3 Norfloxacin R R S R 4 Pefloxacin R R R R 5 cefazolin R R MS R 6 Ceprofloxin S S S S 7 Chloramphenicol S S S S 8 Tobramycin MS MS MS MS 9 Moxifloxacin MS MS MS MS From this study we conclude that antimicrobial susceptibilities in Staph. aureus often differed with regard to sex and age of the humans. The prevalence and degree of antimicrobial resistance in suppurative infections are increasing worldwide (Werckenthinet al., 2001). Staphylococci have shown a frequent and rapid development and spread of antimicrobial resistance. Unfortunately, this development has not been sources (Seguinet al., 1999; Aarestrup and Jensen, 1998)where methicillin resistance is a growing problem (Witte, 1999). However, newer broadspectrum antibiotics, such as enrofloxin, are increasingly being used for the treatment and resistance rates may also increase in staphylococci with their frequent use

4 Surprisingly, tetracycline and chloramphenicol is still an effective antimicrobial in reproductive and abscess sites (Schreiner, 1984). Our findings showed increased resistance to norfloxacin, pefloxacin, and cefazolin. The sex and age of individuals have been suggested as predisposing factors for the development of certain bacterial infections (Picardand Goullet, 1989). Such data might provide useful insights into dynamic aspects of ageand sex related processes involved in the development of antibiotic resistance. Although chloramphenicol and tetracycline resistance iscommonly reported in Staphylococci(Schwarz and Wang, 1993; Schwarz et al., 1998; Alekshun and Levy, 2000),our findings showed sensitive to chloramphenicol. It has even been reported that these antimicrobial agents may be more effective in adults since clinical use in Children is limited due to ageassociated toxicity (Lewis and Reeves, 1994). ACKNOWLEDGEMENT We the authors are cordially thankful to Bejansingh eye hospital, Nagercoil, and the management and principal of Udaya College of Arts and Science, Vellamodi. Also we thank Ms. V.R. Shanthini for her intense help throughout the research work. REFERENCES. Aarestrup, F.M. and Jensen, N.E. Microbiology of Drug Resistance4: ,1998. Alekshun, M.N. and Levy, S.B.In Bacterial Stress Responses pp Washington, DC: ASM Press, Biberstein, E.L., Jang, S.S. and Hirsh, D.C.Journal of Clinical Microbiology19: , Cruickshank R.Medical Microbiology: a guide to diagnosis and control of infection, 11th Edn. E & S Livingstone Ltd. Edinburgh and London; pp 896, Euzeby, J.P.International Journal of Systemic Bacteriology47: , Flournoy, D.J., Murray, C.K. and Vernon, A.N.Methods and Findings in Experimental and Clinical Pharmacology 11: , Flournoy, D.J., Parker, N.S. and Sackett, W.R. Laboratory Medicine10: 39 41, Hoekstra, K.A. and Paulton, R.J.L.Letters in Applied Microbiology22: , Holt, J.G., Krieg, N.R., Sneathm, P.H.A., Staley, J.T. and Williams, S.T. Bergey s Manual of Determinative Bacteriology, 9th edn.baltimore, MD: Williams and Williams, Jorgensen, J.H., Tunridge, J.D. and Washington, J.A. Antibacterial Susceptibility Tests: Dilution and Disk Diffusion Methods.In Manual of Clinical Microbiology, 7th edition eds, Kloos, W.E. Journal of Applied Bacteriology Symposium Suppl.69: 25S 37S, Kloos, W.E. and Schleifer, K.H.Journal of Clinical Microbiology1: 82 88, Lewis, D.A. and Reeves, D.S.Journal of Antimicrobial Chemotherapy34 (Suppl. A): 11 18, Moorthi, C.P., Paul, R., Srinivasan A. and Kumar, C.S.Der Pharmacia Lettre, 3(3): , 2011.

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